Key Laboratory of Molecular Microbiology and Technology, College of Life Sciences, Ministry of Education, Nankai University, 300071, Tianjin, PR China.
Microb Cell Fact. 2023 Aug 27;22(1):162. doi: 10.1186/s12934-023-02159-2.
Poly-β-hydroxybutyrate (PHB), produced by a variety of microbial organisms, is a good substitute for petrochemically derived plastics due to its excellent properties such as biocompatibility and biodegradability. The high cost of PHB production is a huge barrier for application and popularization of such bioplastics. Thus, the reduction of the cost is of great interest. Using low-cost substrates for PHB production is an efficient and feasible means to reduce manufacturing costs, and the construction of microbial cell factories is also a potential way to reduce the cost.
In this study, an engineered Sphingomonas sanxanigenens strain to produce PHB by blocking the biosynthetic pathway of exopolysaccharide was constructed, and the resulting strain was named NXdE. NXdE could produce 9.24 ± 0.11 g/L PHB with a content of 84.0% cell dry weight (CDW) using glucose as a sole carbon source, which was significantly increased by 76.3% compared with the original strain NX02. Subsequently, the PHB yield of NXdE under the co-substrate with different proportions of glucose and xylose was also investigated, and results showed that the addition of xylose would reduce the PHB production. Hence, the Dahms pathway, which directly converted D-xylose into pyruvate in four sequential enzymatic steps, was enhanced by overexpressing the genes xylB, xylC, and kdpgA encoding xylose dehydrogenase, gluconolactonase, and aldolase in different combinations. The final strain NX02 (ΔssB, pBTxylBxylCkdpgA) (named NXdE II) could successfully co-utilize glucose and xylose from corn straw total hydrolysate (CSTH) to produce 21.49 ± 0.67 g/L PHB with a content of 91.2% CDW, representing a 4.10-fold increase compared to the original strain NX02.
The engineered strain NXdE II could co-utilize glucose and xylose from corn straw hydrolysate, and had a significant increase not only in cell growth but also in PHB yield and content. This work provided a new host strain and strategy for utilization of lignocellulosic biomass such as corn straw to produce intracellular products like PHB.
聚-β-羟基丁酸酯(PHB)由各种微生物产生,由于其具有生物相容性和可生物降解性等优良性能,是石油化工塑料的良好替代品。PHB 生产成本高是此类生物塑料应用和推广的巨大障碍。因此,降低成本具有重要意义。使用低成本基质生产 PHB 是降低制造成本的有效可行手段,构建微生物细胞工厂也是降低成本的潜在途径。
本研究构建了一株通过阻断胞外多糖生物合成途径生产 PHB 的工程化泉发单胞菌(Sphingomonas sanxanigenens)菌株,并将其命名为 NXdE。当以葡萄糖为唯一碳源时,NXdE 可产生 9.24±0.11g/L 的 PHB,细胞干重(CDW)中 PHB 的含量为 84.0%,与原始菌株 NX02 相比,产量显著提高了 76.3%。随后,还研究了 NXdE 在不同比例葡萄糖和木糖共底物条件下的 PHB 产率,结果表明木糖的添加会降低 PHB 的产量。因此,通过过表达编码木酮糖脱氢酶、葡萄糖酸内酯酶和醛缩酶的基因 xylB、xylC 和 kdpgA,增强了直接将 D-木糖在四个连续酶步骤中转化为丙酮酸的 Dahms 途径。最终的菌株 NX02(ΔssB,pBTxylBxylCkdpgA)(命名为 NXdE II)可以成功地从玉米秸秆总水解物(CSTH)共利用葡萄糖和木糖生产 21.49±0.67g/L 的 PHB,CDW 中 PHB 的含量为 91.2%,与原始菌株 NX02 相比,产量提高了 4.10 倍。
工程菌株 NXdE II 可以从玉米秸秆水解物中共利用葡萄糖和木糖,不仅细胞生长显著增加,而且 PHB 的产率和含量也显著增加。这项工作为利用玉米秸秆等木质纤维素生物质生产 PHB 等细胞内产物提供了一种新的宿主菌株和策略。